Effect of dietary vitamin E supplement on the reproductive performance and gene expression profiling in Ompok bimaculatus during breeding period.

This study aimed to evaluate the effect of vitamin E on the reproductive responses and the expression of reproductive genes in butter catfish (Ompok bimaculatus). Diets with different levels of vitamin E supplementation (0, 100, and 150 mg/kg) were provided to groups of fish for 60 days. After rearing for 60 days fish were induced with synthetic hormone, that is, Ovatide to study the breeding performance. Ovatide was administrated intramuscularly at the rate of 0.5 and 0.25 mL/kg of body weight for females and males, respectively, in all the treatments. Breeding performance result shows that supplements of vitamin E positively influenced the GSI%, fertilization rate, fecundity, and hatching rate. To understand the relationship between vitamin E and the breeding performance of the fish at the molecular level, a gene expression study was conducted. This study employed real-time PCR for the selected genes critical for reproductive function. In the brain, testis, and ovary dietary vitamin E supplementation of 100 mg/kg significantly enhanced the mRNA transcription of FSHR, Brdt, ESR1, 17beta2, and LHR. Hence, it can be said that 100 mg/kg of vitamin E supplementation in the diet during the reproductive period of O. bimaculatus could improve breeding performance and the mRNA expression of reproductive hormone receptor genes in both males and females of O. bimaculatus.

Unveiling the molecular mechanisms of pigmentation control in Queen Loach, Botia dario (Hamilton, 1822): Insights from sesame seed and marigold-induced antityrosinase effects.

Fish pigmentation study can reveal understandings in dermatological research based on functional genomics. Cultured ornamental fish becomes dull coloured and antityrosinase activity through sesame seed may enhance skin colour, which has not been studied. Botia dario is an indigenous fish, having ornamental and aesthetic value and can be studied as a model for fish pigmentation genetics. In this study, fish specimens were fed with 15% marigold petal meal along with 5, 10 and 15% w/w sesame seed in diet. Pigmentation genes, that is, tyr, tyrp1a, asip1, gnaq, kitlga, mc1r, mitf, pax7a, rab38, slc7a11, sox9a, sox10, csf1r, bcdo2 and gsta2 in skin and immunogens, that is, il20, nramp, tlr9 and trail in kidney were studied. Gene expression in tissues revealed enhanced pigmentation and immunity as well as the role of tyr, tyrp1a and asip1 in pigmentation. Immunogenes and blood parameters confirmed the best pigmentation diet. Colorimetric analysis also showed the enhancement of pigmentation. Insights from sesame seed and marigold-induced antityrosinase effects will be applied in aquaculture to develop natural, dietary formulations that will enhance pigmentation in ornamental fish, leading to improved skin colour and market value.

Stocking density affects immune and stress-related gene expression of Butter catfish (Ompok bimaculatus) fry in biofloc landscapes.

Scientific research into fish wellness is critical, and the concerns about crowding-related stress due to increased stocking density are inevitable. Taking this into consideration, the study defines the physiological signature of Ompok bimaculatus (Butter catfish) in a biofloc system when subjected to varying levels of stocking density. Fish (mean weight = 1.21 g ± 0.08, n = 600) were randomly stocked in 40-L glass aquaria at stocking densities of 0.5 g/L (T1), 1 g/L (T2), 1.5 g/L (T3), and 2 g/L (T4) and fed a 35% protein diet. After the 90-day trial, the physio-biochemical, molecular, and tissue-level changes were assessed. An integrated biomarker response (IBR) analysis for the key stress indicators aided us in better understanding them. There was a significant difference in blood count between T1 and T4 (total erythrocyte count, hemoglobin, and packed cell volume). T1 had higher levels of globulin and total plasma protein, but T2 had higher levels of albumin. Only in T1 did the respiratory burst and lysozyme activity appear to be higher (p < 0.05). Increased stocking densities had a significant impact on the liver function enzymes, GOT and GPT (p < 0.05). In comparison to lower densities (T1 & T2), higher stocking density (T3 & T4) was found to raise glucose and cortisol levels (p < 0.05). Antioxidant enzymes such as catalase, glutathione-S-transferase, and malondialdehyde were found to be more pronounced in lower density tissues (T1). Furthermore, the IBR plots show that lower densities have better health than higher densities. At higher stocking densities, mRNA expression of HSP70, IL-1, and IL-20 increased (p < 0.05) in kidney and liver tissues. The Nrf-2 and Tlr-9 genes were also upregulated. Also, when stocking density was increased, tissue-level histo-architectural changes were more pronounced than when stocking density was kept low. The findings of this study show that the welfare of Butter catfish cultured at high density in biofloc systems suffers from severe stress, and therefore draw more attention to the development of a species-specific standard rearing methodology in the pursuit of a profitable aqua-farming enterprise.

Isolation of Aeromonas salmonicida subspecies salmonicida from aquaculture environment in India: Polyphasic identification, virulence characterization, and antibiotic susceptibility.

This study reports the polyphasic identification, characterization of virulence potential, and antibiotic susceptibility of Aeromonas salmonicida subspecies salmonicida COFCAU_AS, isolated from an aquaculture system in India. The physiological, biochemical, 16s rRNA gene sequencing and PAAS PCR test identified the strain as Aeromonas salmonicida. The MIY PCR tests established the subspecies as 'salmonicida'. The in vitro tests showed the isolated bacterium as haemolytic with casein, lipid, starch, and gelatin hydrolysis activity, indicating its pathogenic attributes. It also showed the ability to produce slime and biofilm, and additionally, it possessed an A-layer surface protein. In vivo pathogenicity test was performed to determine the LD50 dose of the bacterium in Labeo rohita fingerlings (14.42 ± 1.01 g), which was found to be 106.9 cells fish-1. The bacteria-challenged fingerlings showed skin lesions, erythema at the base of the fins, dropsy, and ulcer. Almost identical clinical signs and mortalities were observed when the same LD50 dose was injected into other Indian major carp species, L. catla and Cirrhinus mrigala. Out of the twelve virulent genes screened, the presence of nine genes viz., aerA, act, ast, alt, hlyA, vapA, exsA, fstA, and lip were detected, whereas ascV, ascC, and ela genes were absent. The A. salmonicida subsp. salmonicida COFCAU_AS was resistant to antibiotics such as penicillin G, rifampicin, ampicillin, and vancomycin while highly sensitive to amoxiclav, nalidixic acid, chloramphenicol, ciprofloxacin, and tetracycline. In summary, we have isolated a virulent A. salmonicida subsp. salmonicida from a tropical aquaculture pond which can cause significant mortality and morbidity in Indian major carp species.

Immune-biochemical response and immune gene expression profiling of Labeo rohita fingerlings fed with ethanolic tea leaf extracts and its survivability against Aeromonas hydrophila infection.

The present study was conducted to evaluate the immunostimulatory effect of tea leaf extract (Camellia sinensis) on Labeo rohita and its resistance against Aeromonas hydrophila infection. The ethanolic extract of green tea (GTEE) was found to be the most potent as compared to other solvent extract which was used for further study. It was used to evaluate immune-biochemical response of L. rohita fingerlings, fed with tea leaf extract (control- 0.0%, 0.2% (T1), 0.4% (T2), 0.8% (T3) and 1% (T4) of GTEE kg-1 feed). Different biochemical parameters like glucose, ALP, GPT, GOT, and immunological parameters like lysozyme activity, NBT, anti-protease activity, myeloperoxidase activity, plasma protein, and immune relevant genes (IL-10, C3, Lysozyme G type and iNOS) expressions were carried out. The immunological parameters such as lysozyme activity, NBT and myeloperoxidase activity showed significantly high value once fed with GTEE incorporated diets. Significant up-regulation of immune genes indicated the enhancement of immune response at molecular level. The biochemical parameters were found to be significantly decreasing, indicating that the extract had hepato-protective effect and can help to overcome stress. The fish, fed with GTEE incorporated diets, showed resistance against A. hydrophila when compared with the control group. 0.2% GTEE showed the highest post-challenged survival (76.67%). From the present study, it is concluded that GTEE @ 0.2% can be used as potent immunostimulant as a sustainable alternative prophylactic and therapeutic agent in aquaculture.

Molecular characterization and structural dynamics of Aquaporin1 from walking catfish in lipid bilayers.

Aquaporin's (AQPs) are the major superfamily of small integral membrane proteins that facilitates transportation of water, urea, ammonia, glycerol and ions across biological cell membranes. Despite of recent advancements made in understanding the biology of Aquaporin's, only few isoforms of aquaporin 1 (AQP1) some of the teleost fish species have been characterized at molecular scale. In this study, we made an attempt to elucidate the molecular mechanism of water transportation in AQP1 from walking catfish (Clarias batrachus), a model species capable of breathing in air and inhabits in challenging environments. Using state-of-the-art computational modelling and all-atoms molecular dynamics simulation, we explored the structural dynamics of full-length aquaporin 1 from walking catfish (CbAQP1) in lipid mimetic bilayers. Unlike AQP1 of human and bovine, structural ensembles of CbAQP1 from MD revealed discrete positioning of pore lining residues at the intracellular end. Snapshots from MD simulation displayed differential dynamics of aromatic/arginine (ar/R) filter and extracellular loop C bridging transmembrane (TM) helix H3 and H4. Distinct conformation of large extracellular loops, loop bridging TM2 domain and HB helix along with positioning of selectivity filter lining residues controls the permeability of water across the bilayer. Moreover, the identified unique and conserved lipid binding sites with 100% lipid occupancy signifies lipid mediated structural dynamics of CbAQP1. All-together, this is the first ever report on structural-dynamics of aquaporin 1 in walking catfish which will be useful to understand the molecular basis of transportation of water and other small molecules under varying degree of hyperosmotic environment.

Influence of the extent of enzymatic hydrolysis on the functional properties of protein hydrolysates from visceral waste of Labeo rohita.

The study aimed to evaluate the extent of hydrolysis on the functional properties of fish protein hydrolysates from fish wastes. Fish protein hydrolysate (FPH) was prepared from the visceral waste of Rohu (Labeo rohita) using Alcalase®. Independent variables of the hydrolysis conditions, namely, time, temperature and enzyme-substrate ratio were optimized through response surface methodology using a completely randomized factorial design. Functional properties of FPH, such as antioxidant activity, metal chelating activity, emulsifying and foaming capacity were studied. Pre-heating of the raw viscera resulted 83% reduction of lipid content in the FPH. The DH was found to be significantly influenced by the hydrolysis conditions studied. The gel electrophoretic study showed that FPH contained peptides in the range of < 10-25 kDa. Optimum degree of hydrolysis (DH) was obtained as 34.7% at 87.5 min, 48.64 °C and enzyme concentration of 0.99%. The results demonstrated that the antioxidant and metal chelating properties of FPH increased with the increase of DH. The surface active properties like emulsifying and foaming capacity were found to be decreased with the increased levels of hydrolysis. These results suggest that hydrolysates from visceral waste of Rohu could find potential use as supplement in animal feed.

Genetics of colour variation in wild versus cultured queen loach, Botia dario (Hamilton, 1822).

The Queen loach (Botia dario), an ornamental fish species having export potential, belongs to family Cobitidae of order Cypriniformes. The dull colouration in captive condition as compared to nature is a drawback in ornamental fisheries. We report the first comparative transcriptomic analysis of Cultured (CBD) and Natural (NBD) B. dario using bioinformatics tools. Total 26 and 7 key genes for melanin and carotenoid colouration were found, respectively. KEGG pathway annotations of the genes were carried out, to annotate and describe their relevance for pigmentation. The qPCR validation of genes confirmed their expression pattern in the skin and muscle. Differential expression of, slc7a11, asip1, mc1r, dct, tyrp1a, tyr, bcdo2, csf1r, plin2, gsta2, star3 and stard5 in the skin and muscle tissues revealed the reasons for wild versus cultured colour variation. The molecular data was further supported by low yellowness and redness values of CBD skin and muscle in a colorimeter.

DNA barcoding and phylogenetics of freshwater fish fauna of Ranganadi River, Arunachal Pradesh.

Arunachal Pradesh, the largest state of North-East India covers almost 60.93% of the Eastern Himalayan hotspot. Fish diversity and species identification is utmost important for fisheries management. But, in some cases morphological characteristics based identification is difficult for a non-specialist to perform. In view of the above, the present study emphasized on the assessment of DNA barcoding, phylogenetics and genetic diversity of fish species in the Ranganadi River, Arunachal Pradesh, India. India. Arunachal Pradesh, the largest state of North-East India covers almost 60.93% of the Eastern Himalayan hotspot. Altogether 114 specimens, representing 22 species, belonging to 3 orders and 5 families were successfully barcoded and found to be 98-100% identical from both GenBank and BOLD databases. Out of these 22 fish species, it was found that one species assessed was Endangered, three species as Near Threatened and one species as Vulnerable. A Neighbour Joining (NJ) tree was constructed using Rstudio for the purpose of a phylogenetic analysis of the identified species. The barcoding gap analysis using K2P, P-distance and Jukes-Cantor was done to detect the presence of cryptic species and barcoding success. The nucleotide base composition and genetic distance analysis were also performed, using MEGA 6.0. DNA Sequence Polymorphism v6.12.03 analysis revealed the nucleotide diversity (p) and haplotype diversity (Hd). The Hd for the whole dataset was found to be 0.975, which showed high genetic diversity in the Ranganadi River. Both morphological key identifying characters and molecular data corroborated the phylogenetic analysis. This COI barcode library, generated in the present study, not only helped in species identification and molecular study, but also in cryptic species identification.

Mystus prabini, a new species of catfish (Siluriformes: Bagridae) from Arunachal Pradesh, north-eastern, India.

Mystus prabini, new species, is described from the Sinkin and the Dibang River in the Lower Dibang Valley District of Arunachal Pradesh, India. The new species differs from all South-Asian congeners except M. bleekeri, M. cavasius, M. zeylanicus,  M. falcarius, M. seengtee, M. cineraceus, M. ngasep, M. rufescens and M. ankutta in having a long adipose fin that reaches anteriorly (vs. distinctly does not reach) the base of the last dorsal-fin ray. The new species can be distinguished from the named nine species in having (vs. lacking) a narrow black mid-lateral stripe extending from the anterior region of tympanic spot to the rounded black spot at the caudal-fin base. The analysis of mitochondrial cytochrome c oxidase subunit I (COI) gene sequence shows that the K2P value between Mystus prabini and all other Mystus species ranges from 8.6-22.1%. Mystus prabini is closest genetically to M. bleekeri and M. albolineatus, from which species it has a genetic distance of 8.6% and 13.9%, respectively. The genetic distance (K2P) between the new species and M. dibrugarensis is 21.1%.

Immunity augmentation in Botia dario (Hamilton, 1822) through carotenoid: a dietary approach.

The immunity and health status of ornamental fish is an important aspect, as they are kept in a confined environment and various stressful conditions which lead to depletion of overall colourful appearance and mortality. The carotenoids can act as immunity boosters in captive aquarium system and may be supplemented in the feed as aquarium fish have no access to natural carotenoids. The study aimed to assess the role of carotenoid on the immunity of B. dario. Marigold petal meal is an important source of carotenoids and used in experimental diets. Four immunogenes namely IL20, TLR9, TRAIL, and Nramp in B. dario were characterized and also studied for their relative expression in the kidney after feeding the fish with marigold petal meal supplemented diet. The expression pattern of the genes was compared with the fish of nature. The IL20 and Nramp gene were upregulated significantly (p < 0.05) in the fish of nature as compared to the experimental fish at the 60th day of feeding carotenoid-rich diet. But the TLR9 and TRAIL gene was upregulated significantly (p < 0.05) in experimental fish as compared to nature. The haematological parameters of fish after feeding with the experimental enriched diet for 60 days also confirmed the role of carotenoids in immunity.

Molecular Mechanisms of Natural Carotenoid-based Pigmentation of Queen Loach, Botia dario (Hamilton, 1822) Under Captive Condition.

The genetic basis and expression patterns of key genes are important aspects of study to understand the colouration. This trait differs between wild and domesticated fish which is a matter of research. Botia dario is an indigenous fish, having ornamental and aesthetic value, which shows faded appearance in terms of colour in domesticated condition than wild. In the present study the carotenoid-fed B. dario were examined through incorporation of marigold petal meal in the diets at the rate of 5, 10 and 15% w/w along with wild fish. The carotenoid content of tissues that is skin, muscle and intestine along with intensity of colouration increased in a dose dependant manner of carotenoid in the diet. Important carotenoid-based colouration genes that is csf1r, BCDO2, SR-B1, MLN64, STAR5, GSTA2 and PLIN2 were characterized in the fish, to find out their role in fish pigmentation. The significant difference (p < 0.05) in the expression of these genes in different tissues, when compared among carotenoid-fed domesticated and wild fish, revealed the mechanism responsible for faded colouration and also revealed the means to enhance colour in the fish.

Diagnosis of mitogenome for robust phylogeny: A case of Cypriniformes fish group.

Phylogenetic tree using mitochondrial genes and nuclear genes have long been used for augmenting biological classification and understanding evolutionary processes in different lineage of life. But a basic question still exists for finding the most suitable gene for constructing robust phylogenetic tree. Much of the controversy appears due to monophyletic, paraphyletic and polyphyletic clade making deviations from original taxonomy. In the present study we report the first complete mitochondrial genome (mitogenome) of queen loach, generated through next-generation sequencing methods. The assembled mitogenome is a 16,492 bp circular DNA, comprising of 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a control region. Further in this study we also investigated the suitability of different mitochondrial region for phylogenetic analysis in Cyprinidae and loach group. For this genetic tree were constructed on COI, COII, COIII, 16S rRNA, 12S rRNA, Cyt b, ATPase 6, D-loop, ND1, ND2, ND3, ND4, ND5, and ND6 along with complete mitogenome. The complete mitogenome based phylogenetic tree got inclusive support from available classical taxonomy for these groups. On individual gene basis Cyt b, 12S rRNA, ND2 and ND3 also produced perfect clade at family and subfamily level. For rest of the genes polyphyly were observed for the fishes belonging to same family or subfamily which makes their use questionable for phylogenetic tree construction.

Effect of miconazole nitrate on immunological response and its preventive efficacy in Labeo rohita fingerlings against oomycetes Saprolegnia parasitica.

This study evaluated the effect of sublethal doses of antifungal drug miconazole nitrate (MCZ) on immunological responses and its role as a prophylactic drug against S. parasitica in Labeo rohita fingerlings. Fish were fed with sublethal doses of MCZ, that is, T1-6.30 mgMCZ kgBW-1 , T2-12.61 mgMCZ kgBW-1 and T3-25.22 mgMCZ kgBW-1 , and sampling was done at different time intervals for 240 hr. Immunological parameters viz. lysozyme activity, oxygen radical production and plasma antiprotease activity showed significant enhancement (p < 0.05) in fish fed with T2 and T3 doses. Expression of immune-relevant genes such as TLR-22 and ß2-M showed significantly higher expression at 6 hr and 24 hr of sampling in both liver and head kidney. However, these genes showed a downregulation after 120 hr of sampling in both the tissues. Preventive efficacy study showed that single dose of MCZ provides protection against oomycetes up to the fourth day of infection. Significantly higher mortality was observed in control diet-fed fish as compared to fish fed with MCZ medicated diet. Thus, it can be concluded that the MCZ can act as a potent antifungal agent for preventing oomycetes infection as well as to enhance the immune response.

The cellular localization and redistribution of multiple aquaporin paralogs in the spermatic duct epithelium of a maturing marine teleost.

Aquaporin-mediated fluid transport in the mammalian efferent duct and epididymis is believed to play a role in sperm maturation and concentration. In fish, such as the marine teleost gilthead seabream (Sparus aurata), the control of fluid homeostasis in the spermatic duct seems also to be crucial for male fertility, but no information exists on the expression and distribution of aquaporins. In this study, reverse transcriptase-polymerase chain reaction and immunoblotting analyses, employing available and newly raised paralog-specific antibodies for seabream aquaporins, indicate that up to nine functional aquaporins, Aqp0a, -1aa, -1ab, -3a, -4a, -7, -8bb, -9b and -10b, are expressed in the spermatic duct. Immunolocalization of the channels in the resting spermatic duct reveals that Aqp0a, -1aa, -4a, -7 and -10b are expressed in the monolayered luminal epithelium, Aqp8b and -9b in smooth muscle fibers, and Aqp1ab and -3a in different interstitial lamina cells. In the epithelial cells, Aqp0a and -1aa are localized in the short apical microvilli, and Aqp4a and -10b show apical and basolateral staining, whereas Aqp7 is solely detected in vesicular compartments. Upon spermiation, an elongation of the epithelial cells sterocilia, as well as the folding of the epithelium, is observed. At this stage, single- and double-immunostaining, using two aquaporin paralogs or the Na+ /K+ -ATPase membrane marker, indicate that Aqp1ab, -3a, -7, -8bb and -9b staining remains unchanged, whereas in epithelial cells Aqp1aa translation is supressed, Aqp4a internalizes, and Aqp0a and -10b accumulate in the apical, lateral and basal plasma membrane. These findings uncover a cell type- and region-specific distribution of multiple aquaporins in the piscine spermatic duct, which shares conserved features of the mammalian system. The data therefore suggest that aquaporins may play different roles in the regulation of fluid homeostasis and sperm maturation in the male reproductive tract of fish.

Effect of feeding Lipopolysaccharide as an immunostimulant on immune response and immune gene expression of Labeo bata.

This study investigates the effects of dietary lipopolysaccharide (LPS) as an immunostimulant on hematology, innate immunity, immune gene expression and protection against Edwardsiella tarda on Labeo bata. A basal diet supplemented with 0, 50, 100 and 150mg LPS kg-1diet was fed to the four different groups for 30days. The haematological (total erythrocyte count, total leukocyte count, total serum protein, albumin and globulin), innate immune parameters (respiratory burst, serum lysozyme, myeloperoxidase and serum bactericidal activity), immune gene expression (C3, ß-2 microglobulin, lysozyme g, transferrin, IFN-1, IFN-γ) were monitored at 7th, 15th, 30th day and one day post challenge (DPC) with E. tarda. All the studied haematological, innate immune parameters and expression of immune gene increased significantly (p≤0.05) in LPS fed group in comparison with control. However the group fed 100mgkg-1 LPS in feed showed highest activity on 7th day and 1DPC. The group fed 100mgkg-1 LPS also recorded highest relative percent survivability after challenge with E. tarda. Therefore this study suggests that LPS at 100mgkg-1 could be used as an immunostimulant in feed to enhance the protection of bata during periods of increased disease risk.

Paraprobiotic preparation from Bacillus amyloliquefaciens FPTB16 modulates immune response and immune relevant gene expression in Catla catla (Hamilton, 1822).

The present study evaluated the paraprobiotic effect of heat-inactivated Bacillus amyloliquefaciens FPTB16 on immunological response and immune relevant gene expression in catla (Catla catla). Heat inactivation of viable cells of B. amyloliquefaciens was done at 60 °C for 2 h. For preparation of paraprobiotic supplemented diet, the heat-inactivated bacteria were added to the basal diet (control) at three different inclusion levels i.e., 107, 108 and 109 cells g-1 diet. Fish (25.98 ± 2.57 g) were fed with these diets and various immune responses and immune relevant gene expressions were measured after 4 weeks of feeding. Biochemical parameters were also measured along with the immunological responses. Immunological parameters viz. oxygen radical production, serum lysozyme activity and total serum protein content showed significant enhancement (p < 0.05) in fish fed with 108 and 109 cells g-1 diet. Significant enhancement in myeloperoxidase activity was observed in all the dietary groups compared to control. Alkaline phosphatase activity showed significant enhancement (p < 0.05) in fish fed with 107 and 108 cells g-1 diet. Biochemical parameters viz. GPT, GOT and glucose content did not show any significant difference in any of the dietary groups. Immune relevant genes viz. IL-1ß, TNF-α, C3 and iNOS showed significantly higher expression in either liver or head-kidney tissues in most of the cases. However, IFN-γ expression showed a down-regulation pattern in both the tissues. In conclusion, the dietary supplementation of heat-inactivated B. amyloliquefaciens enhanced the immunity of catla, particularly at 108 cells g-1 diet. The results collectively suggest the paraprobiotic applicability of B. amyloliquefaciens in aquaculture.

Dual estrogenic regulation of the nuclear progestin receptor and spermatogonial renewal during gilthead seabream (Sparus aurata) spermatogenesis.

Studies in teleosts suggest that progestins have crucial functions during early spermatogenesis. However, the role of the different progestin receptors in these mechanisms is poorly understood. In this work, we investigated the expression pattern and hormonal regulation of the classical nuclear progestin receptor (Pgr) in the gilthead seabream at three different stages of spermatogenesis: the resting (postspawning) phase, onset of spermatogenesis, and spermiation. Immunolocalization experiments using a seabream specific Pgr antibody revealed that the receptor was expressed in Sertoli and Leydig cells, and also in a subset of spermatogonia type A, throughout spermatogenesis. Short-term treatment of testis explants with 17ß-estradiol (E2) increased pgr mRNA expression at all stages, while the progestin 17α,20ß-dihydroxy-4-pregnen-3-one (17,20ßP) had the opposite effect. At the resting stage, Sertoli cell Pgr expression was positively correlated with the occurrence of proliferating spermatogonia type A in the tubules, and both processes were incremented in vitro by E2 likely through the estrogen receptor alpha (Era) expressed in Sertoli and Leydig cells. In contrast, treatment with 17,20ßP downregulated Pgr expression in somatic cells. The androgen 11-ketotestosterone (11-KT) upregulated pgr expression in Leydig cells and promoted the proliferation of mostly spermatogonia type B, but only during spermiation. No relationship between the changes in the cell type-specific expression of the Pgr with the entry into meiosis of germ cells was found. These data suggest a differential steroid regulation of Pgr expression during seabream spermatogenesis and the potential interplay of the E2/Era and 17,20ßP/Pgr pathways for the maintenance of spermatogonial renewal rather than entry into meiosis.

Genetic diversity and demographic history of the giant river catfish Sperata seenghala inferred from mitochondrial DNA markers.

The giant river catfish Sperata seenghala has huge demand in South Asian countries due to its low number of intramuscular bones and nutritive value. However, the culture practises for this fish have not been standardized and the current demand for this fish is being met by capture fisheries only. Unregulated and indiscriminate fishing would lead overexploitation of fish stocks subsequently stock depletion. Genetic diversity between populations would give insight about population structure and demography. In the present study, S. seenghala stocks from three rivers, namely Ganga, Brahmaputra and Mahanadi were characterized using cytochrome b gene and D-loop region. Moderate to high haplotype diversity and low nucleotide diversity values were observed in all populations. Analysis of molecular variance and pairwise FST values showed significant genetic differentiation among populations. Patterns of diversity, haplotype networks and mismatch distribution strongly suggest a historical influence on the genetic structure of S. seenghala populations. S. seenghala stocks from these three rivers are genetically distinct units and management measures should be formulated separately for each population.

Gel properties of sutchi catfish (Pangasius hypophthalmus) surimi as affected by selected washing process and number of washing cycles.

The gel properties of sutchi catfish surimi prepared by conventional washing and alkaline-saline washing method were studied for four washing cycles. Decrease (p < 0.05) in myoglobin content was found in alkaline washing process compared to conventional washing at each washing cycle. The highest hardness, breaking force and deformation was observed in gels prepared from alkaline-saline washing method. Whiteness in conventional washed surimi gels increased non-significantly (p < 0.05) compared to alkaline-saline-washed surimi gels. Protein bands on Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that the protein extractability was more in alkaline-saline washing and disappearance of bands in conventional washing method was observed between 116 and 45 kDa indicating less yield of protein.